GSM7119480: Pool of 3 B16/F10-BrM brains; Mus musculus; RNA-Seq
Mouse brains were processed with the Adult Brain Dissociation Kit (Miltenyi, ref. 130-107-677) using gentleMACS C Tubes (Miltenyi, ref. 130-093-237) and the gentleMACS™ Octo Dissociator (Miltenyi, ref.  130-096-427). For myelin removal, a 30% percoll gradient was used. To isolate the highest possible amount of astrocytes, Anti-ACSA-2 (Miltenyi, ref. 130-097-678) microbeads were used and for dead cell removal, Debris Removal solution (Miltenyi, ref. 130-109-398) was added prior to analysis. ~10,000 cells per condition were loaded onto a 10x Chromium Single Cell Controller chip B (10x Genomics) as described by the manufacturer (Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3, PN-1000075). Generation of gel beads in emulsion (GEMs), barcoding, GEM-RT clean-up, cDNA amplification and library construction were all performed as recommended in the manufacturer's protocol.